Research
Because the 5' → 3' exonuclease activity of DNA polymerase I from E. coli makes it unsuitable for many applications, the Klenow fragment, which lacks this activity, can be very useful in research. The Klenow fragment is extremely useful for research-based tasks such as:
- Synthesis of double-stranded DNA from single-stranded templates
- Filling in (meaning removal of overhangs to create blunt ends) recessed 3' ends of DNA fragments
- Digesting away protruding 3' overhangs
- Preparation of radioactive DNA probes
The Klenow fragment was also the original enzyme used for greatly amplifying segments of DNA in the polymerase chain reaction (PCR) process, before being replaced by thermostable enzymes such as Taq polymerase.
Read more about this topic: Klenow Fragment
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